Project Summary The University of Nebraska Medical Center (UNMC) requests funds to purchase an FEI Apreo VS? serial blockface imaging scanning electron microscope (SBFSEM) to provide state-of-the-art imaging capabilities that are not currently available at UNMC or in the Omaha metropolitan region. The Apreo VS? is a highly innovative electron microscope that combines the imaging power of a field emission electron gun scanning electron microscope with precision ultramicrotomy, multi-energy deconvolution, novel integrated detectors, and powerful image analysis software to enable 3D visualization and quantitative analysis at nanometer resolution. SBFSEM has advantages over other methods of 3D electron microscopy in that both operation of the microscope and image collection/alignment are automated processes, which reduces skill requirements, increases reproducibility, and enhances accessibility for the average investigator. Quantitative 3D imaging will enable detailed studies of cell and tissue ultrastructure and will improve quantification of number, volume, surface area and morphology of organelles and cell structures for diverse medically relevant projects. The proposed project is viable because of strong investigator support illustrated by the 16 NIH-funded and other projects that include studies of cancer biology, neurosensory systems development and disease, biofilms and infectious disease, immunology, cardiac and vascular disease, nanopharmaceuticals, and tissue engineering, and involve investigators from several academic divisions at UNMC,as well as from nearby institutions,such as University of Nebraska-Lincoln, Creighton University, and Boys Town National Research Hospital. Each project is designed to take full advantage of SBFSEM to advance knowledge of physiological and disease echanisms, or to enable development of novel pharmaceuticals or bioengineering approaches for regenerative medicine. These projects enjoy strong technical support that includes two MSA-certified Electron Microscopy Specialists, one with nearly forty years of experience in transmission and scanning electron microscopy; the Director of the Research Information Technology Office at UNMC (Dr. Mudgapalli), and a 3D visualization and image segmentation expert with experience in Amira software (Mr. Glass). The proposal also has strong institutional and inter-institutional support as evidenced by salary and service agreement support by the Vice Chancellor of Research (Dr. Larsen), as well as letters of support from the Monroe-Meyer Institute, the Nebraska Center for Cell Signaling (COBRE funded), and Boys Town National Research Hospital (Omaha). The Apreo VS? will be housed in and administered by the Electron Microscopy Core Facility (EMCF), which has served UNMC and nearby institutions for four decades and is currently the only full service biological research-oriented electron microscopy facility in the metropolitan area. The combination of strong user interest, diverse and significant projects, technical expertise, administrative experience, and a solid long-term plan will ensure successful implementation and extensive use of the Apreo VS?.